The broad goals of this project are to understand the mechanisms of action of basic amino acid transport systems, particularly of those of arginine, lysine, ornithine and histidine in Escherichia coli. Special emphasis will be placed upon the characterization of permease defective mutants obtained by localized mutagenesis procedure. The knowledge obtained by transport and genetic studies on these mutants will be correlated with physical and chemical studies on osmotic shock released binding proteins involved in the transport of basic amino acids. Recently a procedure has been developed for the obtention of a unit of two fused episomes carrying the attachment site for phage phi 80 and the arginine permease locus (argP) from wild type Escherichia coli. After the attainment of transducing phage particles carrying basic amino acid permease genes from wild type Escherichia coli as well as from permease defective mutants, the amplified synthesis of permease gene products will be attempted. Efforts will also be made to obtain permease gene products as soluble proteins by using lysis defective phage mutants. Subsequent to physical and chemical studies of the different permease components, attempts would be made to reconstitute transport systems either by forming membrane structures with soluble proteins and lipids or by replacing the permeability components to membrane preparation from permease deficient cells.